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Nucleic Acids Research, 1980, Vol. 8, No. 19 4485-4500
© 1980


MOLECULAR BIOLOGY

Fine structure mapping of an avian tumor virus RNA by immunoelectron microscopy

H. Castleman, R.D. Meredith and B.F. Erlanger

Department of Microbiology, Columbia University, Cancer Center/Institute of Cancer Research New York, NY 10032, USA

Received June 20, 1980. The RNA of a deleted strain (lacking Src gene) of an avian sarcoma virus (ASV) was examined by a newly developed immunoelectron microscopic procedure which uses anti-nucleotide antibodies as probes. After denaturation of the RNA and reaction with a high affinity, highly specific anti-7-methylguanosine-5'-phosphate (anti-pm 7G), 81% of 106 molecules examined were found to have antibody at one terminus, in agreement with the presence of a pm 7G cap in ASV-RNA. Hapten inhibition by pm 7G could be demonstrated. Experiments with anti-A and with anti-poly A gave results consistent with the known structure of ASV-RNA, in particular the presence of a 3' poly A tail. These studies illustrate the feasibility of using anti-nucleotide antibodies in a combined immunochemical and electron microscopic study of the fine structure of nucleic acids.


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